新基因pp3774抑制NIH/3T3细胞生长机制初探  

Preliminary study of mechanism on a novel gene pp3774 inhibiting NIH/3T3 cells growth

作　　者：张锋锐[1] 李锦军[1] 王春艳[1] 朱丽[1] 徐文[1] 顾健人[1] 

机构地区：[1]上海交通大学肿瘤研究所上海市肿瘤研究所癌基因及相关基因国家重点实验室,上海200032

出　　处：《肿瘤》2005年第6期550-554,共5页Tumor

基　　金：国家863项目(编号:2005AA220040;2004AA227090);国家十五攻关重大项目(编号2003BA711A02-1);上海市科委重大基础研究课题(编号:03DJ14007);上海市卫生局科技发展基金(编号:034012)

摘　　要：目的探讨新基因pp3774在NIH／3T3细胞生长调节中的可能机制。方法用pp3774-HA融合表达质粒通过脂质体法转染NIH／3T3细胞,G418筛选,建立稳定细胞系,并用RT-PCR检测pp3774在NIH／3T3细胞中mRNA的表达状况；利用Atlas cDNA array分析pp3774的异位表达对NIH／3T3细胞基因表达谱的影响；用RT-PCR方法验证Atlas cDNA array杂交结果。结果 pp3774超表达可以明显下调cyclin D1、cyclin E、转录因子Dpl等基因的表达,同时可上调P13K p85α、FGF7、MMP-2等基因的表达。上述基因表达的改变可能通过调控细胞周期实现pp3774对NIH／3T3细胞的生长抑制。结论新基因pp3774抑制NIH／3T3细胞生长可能与pp3774基因下调cyclin D1、cyclin E、转录因子Dpl的表达有关。Objective To explore the possible mechanism of a novel gene pp3774 in NIH/3T3 cell growth regulation. Methods Lipofectamine method was used to transfect pp3774-HA plasmid into NIH/3T3 cell, screened with G418, and stable-expressing cell line was established;RT-PCR was subjected to detect the expression of pp3774 mRNA; Atlas eDNA array was used to analyse the effects of ectopic expression of pp3774 gene on NIH/3T3 genomic profile,which was confirmed with RT-PCR. Results Down-regulation of cyclin D1, cyclin E, transcription factor Dp1 expression, and up-regulation of phosphatidylinositol 3-kinase regulatory subunit, polypeptide 1 （p85a）, growth hormone receptor, fibroblast growth factor 7, matrix metalloproteinase 2 expression were induced by ectopic expression of pp3774. These changes might result in the cell growth inhibition of pp3774 gene in NIH/3T3 cells through cell cycle regulation. Conclusion The inhibition of cell growth in NIH/3T3 may be associated with down-regulation of cyclin D1 ,cyclin E and transcription factor Dp1 by novel gene pp3774.

关 键 词：基因 pp3774 基因表达调控 肿瘤 寡核苷酸排列序列分析 逆转录酶聚合酶链反应 NIH/3T3细胞 

分 类 号：R730.1[医药卫生—肿瘤]